alexa The Determination of Factors involved in Column-Based N
ISSN: 2155-9821

Journal of Bioprocessing & Biotechniques
Open Access

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Research Article

The Determination of Factors involved in Column-Based Nucleic Acid Extraction and Purification

Jun-Jie Poh1,3 and Samuel Ken-En Gan1,2,3*
1 Bioinformatics Institute, Agency for Science, Technology, and Research (A*STAR), 138671, Singapore
2 p53 Laboratory, Agency for Science, Technology, and Research (A*STAR), 138648, Singapore
3 Quintech Life Sciences Pte Ltd, 619933, Singapore
Corresponding Author : Samuel Ken-En Gan
Bioinformatics Institute
Agency for Science, Technology and Research (A*STAR)
30 Biopolis Street, #07-01 Matrix
138671, Singapore
Tel: 65-6478-8417, 65-6407-0584
Fax: 65-6478-9047
E-mail: [email protected]
Received April 25, 2014; Accepted June 06, 2014; Published June 12, 2014
Citation: Poh JJ, Gan SKE (2014) The Determination of Factors Involved in Column-Based Nucleic Acid Extraction and Purification. J Bioprocess Biotech 4:157 doi: 10.4172/2155-9821.1000157
Copyright: © 2014 Poh JJ et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
 

Abstract

DNA extraction methods such as plasmid minipreps, gel, and PCR purifications, are indispensable techniques for genetic manipulations. There are numerous factors that contribute to the efficiency of these processes, which determine the success of complex downstream molecular analytics and diagnostic tests. To study and optimize these factors, we compared our own proprietary buffers to commercially available column-based kits, utilizing their spin columns and protocols. Through systematic substitution of the buffers in the kits with our own proprietary buffers, we selected the highest DNA yielding buffer recipes. Further analysis of the differences between the buffers showed that high concentrations and presence of certain chaotropic agents and cations are necessary for good plasmid miniprep, gel extraction, and PCR purification kits.

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